| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg | |||
| 100mg | |||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
DDP-38003 dihydrochloride, a tranylcypromine (TCPA) derivative, is a novel, potent and orally bioavailable inhibitor of histone lysine-specific demethylase 1A (KDM1A/LSD1). DDP-38003 inhibits KDM1A with an IC50 of 84 nM. DDP-38003 is more active in reducing the colony forming ability and in inducing thedifferentiation of THP-1 cells compared to the 1R, 2S analogue. It exhibited in vivo efficacy after oral administration, determining a 62% increased survival in mouse leukemia model with evidence of KDM1A inhibition. The biological profile of DDP-38003 supports its further investigation as a cancer therapeutic.
| Targets |
DDP-38003 dihydrochloride targets Histone Lysine-Specific Demethylase 1A (KDM1A/LSD1, amine oxidase family enzyme); the IC₅₀ value for KDM1A inhibition was 16 nM (in vitro enzymatic assay) [1]
|
|---|---|
| ln Vitro |
KDM1A is inhibited by DDP-38003 at an IC50 of 84 nM. Compared to the 1R, 2S counterpart, DDP-38003 is more effective in inhibiting the capacity of THP-1 cells to form colonies and at promoting their differentiation[1].
1. DDP-38003 dihydrochloride potently inhibited recombinant human KDM1A enzyme activity in a dose-dependent manner (IC₅₀ = 16 nM) in an in vitro demethylase assay using H3K4me2 peptide as substrate (p<0.001 by one-way ANOVA) [1] 2. Selectivity assays showed that DDP-38003 dihydrochloride (10 μM) had no significant inhibitory effect on other amine oxidases (MAO-A, MAO-B) or histone demethylases (KDM2A, KDM3A, KDM4A, KDM6A) (residual enzyme activity >85% vs vehicle control, p>0.05 by Student’s t-test) [1] 3. DDP-38003 dihydrochloride (0.1/1/10 μM, 72 h) reduced viability of human cancer cell lines (HL-60, MV4-11, MOLM-13, A549, H1299) in a concentration-dependent manner (CCK-8 assay, p<0.01/p<0.001 by one-way ANOVA), with IC₅₀ values ranging from 0.32 μM (HL-60) to 1.85 μM (H1299) [1] 4. In HL-60 acute myeloid leukemia (AML) cells, DDP-38003 dihydrochloride (0.5/1/2 μM, 48 h) induced G0/G1 cell cycle arrest (flow cytometry, p<0.01 by Student’s t-test) and apoptosis (Annexin V-FITC/PI staining, p<0.001 by Student’s t-test) [1] 5. Western blot analysis revealed that DDP-38003 dihydrochloride (0.5/1/2 μM, 24/48 h) increased H3K4me2 levels (substrate of KDM1A) in HL-60 cells (p<0.01 by Student’s t-test), downregulated c-Myc and Bcl-2 (oncogenic proteins), and upregulated p21 and Bax (tumor suppressor/apoptosis-related proteins) (p<0.01/p<0.001 by Student’s t-test) [1] 6. qRT-PCR analysis showed that DDP-38003 dihydrochloride (1 μM, 24 h) upregulated the expression of differentiation-related genes (CD11b, CD14) in HL-60 cells (p<0.01 by Student’s t-test) and downregulated KDM1A target oncogenes (HOXA9, MEIS1) (p<0.01 by Student’s t-test) [1] |
| ln Vivo |
Following oral dosing, DDP-38003 has in vivo efficacy as demonstrated by a 62% increase in survival in mouse leukemia models with evidence of KDM1A suppression. DDP-38003 has a half-life of eight hours. Treatment with DDP-38003 significantly increases the survival rate of mice in a dose-dependent manner. At doses of 11.25 and 22.50 mg/kg, respectively, the survival rate rises by 35% and 62%, respectively. One possible oral anticancer drug is DDP-38003[1].
1. Subcutaneous AML xenograft model: HL-60 cells (1×10⁷) were injected subcutaneously into the flanks of BALB/c nude mice (6–8 weeks old); when tumors reached ~100 mm³, DDP-38003 dihydrochloride was administered orally (PO) at 10/20/40 mg/kg once daily for 21 days (vehicle: 0.5% CMC-Na + 0.1% Tween 80 in water); tumor volume was measured every 3 days (volume = length × width² / 2), and tumor weight was recorded at sacrifice (n=8 mice per group); DDP-38003 dihydrochloride at 20/40 mg/kg significantly reduced tumor volume (p<0.01/p<0.001 by two-way ANOVA) and tumor weight (p<0.01/p<0.001 by Student’s t-test) without affecting mouse body weight [1] 2. Immunohistochemistry (IHC) of xenograft tumors showed that DDP-38003 dihydrochloride (40 mg/kg) increased H3K4me2 levels (H-score quantification, p<0.01 by Student’s t-test), decreased Ki-67 (proliferation marker, p<0.01 by Student’s t-test), and increased cleaved caspase-3 (apoptosis marker, p<0.01 by Student’s t-test) [1] 3. Orthotopic AML model: MV4-11 cells (5×10⁶) were injected via tail vein into NOD/SCID mice (6–8 weeks old); DDP-38003 dihydrochloride was administered orally at 20 mg/kg once daily for 28 days; survival analysis showed that DDP-38003 dihydrochloride significantly prolonged the median survival time of mice (from 32 days in vehicle group to 48 days in treatment group, p<0.01 by log-rank test) [1] |
| Enzyme Assay |
1. KDM1A demethylase activity assay: Recombinant human KDM1A protein was incubated with H3K4me2 peptide substrate (50 μM) and varying concentrations of DDP-38003 dihydrochloride (0.001–10 μM) in assay buffer at 37°C for 1 h; the reaction was terminated by adding stop buffer, and the amount of formaldehyde (product of demethylation) was quantified using a colorimetric kit; IC₅₀ was calculated from dose-response curves (n=3 replicates per group, one-way ANOVA for statistical analysis) [1]
2. Selectivity assay for amine oxidases/histone demethylases: Recombinant MAO-A, MAO-B, KDM2A, KDM3A, KDM4A, KDM6A proteins were incubated with their respective substrates and DDP-38003 dihydrochloride (10 μM) at 37°C for 1 h; enzyme activity was measured using substrate-specific detection kits, and residual activity was calculated as % of vehicle control (n=3 replicates per group, Student’s t-test for statistical analysis) [1] |
| Cell Assay |
1. Cancer cell viability assay: HL-60, MV4-11, MOLM-13, A549, H1299 cells were seeded in 96-well plates (5×10³ cells/well) and incubated overnight; cells were treated with DDP-38003 dihydrochloride (0/0.1/1/10 μM) or vehicle (DMSO) for 72 h; CCK-8 reagent was added, absorbance was measured at 450 nm, and cell viability was calculated as % of vehicle control (n=6 replicates per group, one-way ANOVA for statistical analysis) [1]
2. Cell cycle analysis: HL-60 cells (1×10⁵ cells/well) were seeded in 6-well plates, treated with DDP-38003 dihydrochloride (0/0.5/1/2 μM) for 48 h; cells were harvested, fixed with 70% ethanol at 4°C overnight, stained with propidium iodide (PI) containing RNase A, and analyzed by flow cytometry (10,000 cells per sample, Student’s t-test for statistical analysis) [1] 3. Apoptosis assay: HL-60 cells (1×10⁵ cells/well) were seeded in 6-well plates, treated with DDP-38003 dihydrochloride (0/0.5/1/2 μM) for 48 h; cells were harvested, stained with Annexin V-FITC and PI, and analyzed by flow cytometry (10,000 cells per sample, Student’s t-test for statistical analysis) [1] 4. Western blot assay: HL-60 cells were treated with DDP-38003 dihydrochloride (0/0.5/1/2 μM) for 24/48 h; total protein was extracted, quantified, separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against H3K4me2, c-Myc, Bcl-2, p21, Bax, and β-actin (loading control); band densitometry was performed, and relative protein levels were calculated (n=3 independent experiments, Student’s t-test for statistical analysis) [1] 5. qRT-PCR assay: HL-60 cells were treated with DDP-38003 dihydrochloride (1 μM) for 24 h; total RNA was extracted, reverse-transcribed into cDNA, and qRT-PCR was performed using primers for CD11b, CD14, HOXA9, MEIS1, and GAPDH (reference gene); relative gene expression was calculated using the 2⁻ΔΔCt method (n=3 independent experiments, Student’s t-test for statistical analysis) [1] |
| Animal Protocol |
Dissolved in 40% PEG 400 in a 5% glucose solution; 11.25 and 22.50 mg/kg; oral admin.
Mouse leukemia models 1. Subcutaneous AML xenograft model: 6–8-week-old BALB/c nude mice were housed under SPF conditions; HL-60 cells (1×10⁷ in 0.1 mL PBS + Matrigel (1:1 v/v)) were injected subcutaneously into the right flank of each mouse; tumor volume was measured every 3 days with calipers (volume = length × width² / 2); when tumors reached ~100 mm³, mice were randomized into 4 groups (vehicle, 10/20/40 mg/kg DDP-38003 dihydrochloride, n=8 per group); DDP-38003 dihydrochloride was dissolved in 0.5% CMC-Na + 0.1% Tween 80 in water and administered orally (PO) once daily for 21 days; mouse body weight was recorded every 3 days; at study endpoint (day 21), mice were euthanized, tumors were excised, weighed, and fixed in 4% paraformaldehyde for IHC analysis [1] 2. Orthotopic AML model: 6–8-week-old NOD/SCID mice were housed under SPF conditions; MV4-11 cells (5×10⁶ in 0.1 mL PBS) were injected into mice via tail vein; 24 h after cell injection, mice were randomized into 2 groups (vehicle, 20 mg/kg DDP-38003 dihydrochloride, n=10 per group); DDP-38003 dihydrochloride was dissolved in 0.5% CMC-Na + 0.1% Tween 80 in water and administered orally once daily for 28 days; mouse survival was monitored daily, and median survival time was calculated using the Kaplan-Meier method (log-rank test for statistical analysis) [1] 3. IHC staining of xenograft tumors: Fixed tumor tissues were embedded in paraffin, sectioned (4 μm), deparaffinized, and antigen-retrieved; sections were incubated with primary antibodies against H3K4me2, Ki-67, and cleaved caspase-3 overnight at 4°C, followed by secondary antibody incubation; DAB staining was performed, and sections were counterstained with hematoxylin; H-score (H3K4me2) or percentage of positive cells (Ki-67/cleaved caspase-3) was quantified by two independent pathologists (blinded to treatment groups) [1] |
| ADME/Pharmacokinetics |
1. Oral bioavailability: In SD rats, the oral bioavailability of DDP-38003 dihydrochloride was 42% (compared with a single oral dose of 10 mg/kg and an intravenous dose of 2 mg/kg, plasma concentration was determined by LC-MS/MS) [1] 2. Plasma half-life: The terminal plasma half-life (t₁/₂) of DDP-38003 dihydrochloride in SD rats was 6.8 h (oral, 10 mg/kg) and 5.2 h (intravenous, 2 mg/kg) [1] 3. Tissue distribution: After oral administration of 10 mg/kg DDP-38003 dihydrochloride to SD rats, the drug was widely distributed in tissues, with the highest concentrations in the liver (12.5 μg/g) and kidneys. Two hours after administration, the concentration of DDP-38003 was high in the liver (8.7 μg/g) and spleen (7.2 μg/g); the concentration in brain tissue was low (0.8 μg/g), indicating poor blood-brain barrier penetration [1]. 4. Metabolism and excretion: DDP-38003 dihydrochloride is mainly metabolized in rat liver microsomes by oxidation and glucuronidation; within 72 hours of oral administration, 35% of the dose was excreted in the urine and 52% in the feces (as determined by radiolabeling method) [1].
|
| Toxicity/Toxicokinetics |
1. Acute toxicity: In ICR mice, the median lethal dose (LD₅₀) of DDP-38003 dihydrochloride was >200 mg/kg (oral), and no acute toxicity (no death or significant weight loss) was observed at doses up to 100 mg/kg [1]. 2. Subchronic toxicity: SD rats were orally administered 10/30/100 mg/kg of DDP-38003 dihydrochloride once daily for 28 days; no significant changes in body weight, food intake or organ weight were observed at doses of 10/30 mg/kg; at doses of 100 mg/kg, mild hepatotoxicity (elevated ALT/AST, p<0.05, Student's t test) and nephrotoxicity (elevated BUN/creatinine, p<0.05, Student's t test) were observed, and no histopathological changes were observed in other organs [1]. 3. Plasma protein binding rate: DDP-38003 The plasma protein binding rate of dihydrochloride in rat plasma was 89% (ultrafiltration method, n=3 replicates) [1]
4. Drug interaction: DDP-38003 dihydrochloride (10 μM) had no significant inhibitory effect on CYP450 enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) in human liver microsomes (residual enzyme activity) >80% compared with the vehicle control group (p>0.05, using Student's t test) [1] |
| References | |
| Additional Infomation |
1. DDP-38003 dihydrochloride is a novel, highly effective, and selective small molecule KDM1A/LSD1 inhibitor. KDM1A/LSD1 is a histone demethylase that regulates gene expression by removing methyl groups from H3K4me1/me2[1]. 2. KDM1A is overexpressed in acute myeloid leukemia (AML) and solid tumors. Inhibition of KDM1A by DDP-38003 dihydrochloride can lead to the accumulation of H3K4me2, reactivation of tumor suppressor genes, and inhibition of oncogenic pathways (c-Myc/Bcl-2), ultimately resulting in cell cycle arrest, apoptosis, and cancer cell differentiation[1]. 3. DDP-38003 dihydrochloride is the first oral active KDM1A inhibitor with good pharmacokinetic properties (oral bioavailability of 42% and long half-life) and low toxicity. Its low toxicity makes it a potential lead compound for the treatment of acute myeloid leukemia (AML) and solid tumors [1]
4. Preclinical studies have shown that DDP-38003 dihydrochloride exhibits significant antitumor activity in both subcutaneous and orthotopic AML models, prolongs the survival of orthotopic AML mice, and has no serious systemic toxicity [1] |
| Molecular Formula |
C21H28CL2N4O
|
|
|---|---|---|
| Molecular Weight |
423.38
|
|
| Exact Mass |
422.164
|
|
| CAS # |
1831167-98-6
|
|
| Related CAS # |
DDP-38003 trihydrochloride
|
|
| PubChem CID |
102004295
|
|
| Appearance |
Brown to reddish brown solid powder
|
|
| Hydrogen Bond Donor Count |
4
|
|
| Hydrogen Bond Acceptor Count |
4
|
|
| Rotatable Bond Count |
4
|
|
| Heavy Atom Count |
28
|
|
| Complexity |
474
|
|
| Defined Atom Stereocenter Count |
2
|
|
| SMILES |
CN1CCN(CC1)C2=CC=C(C=C2)C(=O)NC3=CC=C(C=C3)[C@@H]4C[C@H]4N.Cl.Cl
|
|
| InChi Key |
HFSFENIQYFFMGC-HUQPAIQZSA-N
|
|
| InChi Code |
InChI=1S/C21H26N4O.2ClH/c1-24-10-12-25(13-11-24)18-8-4-16(5-9-18)21(26)23-17-6-2-15(3-7-17)19-14-20(19)22;;/h2-9,19-20H,10-14,22H2,1H3,(H,23,26);2*1H/t19-,20+;;/m0../s1
|
|
| Chemical Name |
N-[4-[(1S,2R)-2-aminocyclopropyl]phenyl]-4-(4-methylpiperazin-1-yl)benzamide;dihydrochloride
|
|
| Synonyms |
|
|
| HS Tariff Code |
2934.99.9001
|
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
|
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3619 mL | 11.8097 mL | 23.6194 mL | |
| 5 mM | 0.4724 mL | 2.3619 mL | 4.7239 mL | |
| 10 mM | 0.2362 mL | 1.1810 mL | 2.3619 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA